Frontiers in Immunology,
Год журнала:
2025,
Номер
16
Опубликована: Фев. 25, 2025
Antigen
specific
humoral
immunity
can
be
characterized
by
the
analysis
of
serum
antibodies.
While
serological
assays
for
measurement
antibody
levels
are
available,
these
not
quantitative
in
biochemical
sense.
Yet,
understanding
immune
responses
quantitatively
on
systemic
level
would
need
a
universal,
complete,
quantitative,
comparable
method
antigen
antibodies
selected
immunoglobulin
classes.
Here
we
describe
fluorescent,
dual-titration
immunoassay,
which
provides
parameters
that
both
necessary
and
sufficient
to
characterize
response.
For
validation
theory,
used
recombinant
receptor
binding
domain
SARS-CoV-2
as
microspot
arrays
varied
concentration
from
infected
persons
obtain
matrix
data.
Both
titration
curves
were
simultaneously
fitted
using
an
algorithm
based
generalized
logistic
function
adapted
analyzing
variables
binding.
We
obtained
equilibrium
affinity
constants
concentrations
distinct
These
reflect
quality
effective
quantity
antibodies,
respectively.
The
proposed
fluorescent
immunoassay
generate
truly
data
is
suitable
immunological,
medical
systems
biological
analysis.
Journal of Proteome Research,
Год журнала:
2021,
Номер
20(12), С. 5241 - 5263
Опубликована: Окт. 21, 2021
The
study
of
proteins
circulating
in
blood
offers
tremendous
opportunities
to
diagnose,
stratify,
or
possibly
prevent
diseases.
With
recent
technological
advances
and
the
urgent
need
understand
effects
COVID-19,
proteomic
analysis
blood-derived
serum
plasma
has
become
even
more
important
for
studying
human
biology
pathophysiology.
Here
we
provide
views
perspectives
about
developments
possible
clinical
applications
that
use
mass-spectrometry(MS)-
affinity-based
methods.
We
discuss
examples
where
proteomics
contributed
valuable
insights
into
SARS-CoV-2
infections,
aging,
hemostasis
offered
by
combining
with
genetic
data.
As
a
contribution
Human
Proteome
Organization
(HUPO)
Plasma
Project
(HPPP),
present
PeptideAtlas
build
2021-07
comprises
4395
canonical
1482
additional
nonredundant
detected
240
MS-based
experiments.
In
addition,
report
new
Extracellular
Vesicle
2021-06,
which
five
studies
2757
extracellular
vesicles
blood,
74%
(2047)
are
common
PeptideAtlas.
Our
overview
summarizes
advances,
impactful
applications,
ongoing
challenges
translating
utility
precision
medicine.
Science Translational Medicine,
Год журнала:
2022,
Номер
14(655)
Опубликована: Июль 27, 2022
Several
variants
of
severe
acute
respiratory
syndrome
coronavirus
2
(SARS-CoV-2)
have
emerged
during
the
current
disease
2019
(COVID-19)
pandemic.
Although
antibody
cross-reactivity
with
spike
glycoproteins
(S)
diverse
coronaviruses,
including
endemic
common
cold
coronaviruses
(HCoVs),
has
been
documented,
it
remains
unclear
whether
such
responses,
typically
targeting
conserved
S2
subunit,
contribute
to
protection
when
induced
by
infection
or
through
vaccination.
Using
a
mouse
model,
we
found
that
prior
HCoV-OC43
S–targeted
immunity
primes
neutralizing
responses
otherwise
subimmunogenic
SARS-CoV-2
S
exposure
and
promotes
S2-targeting
responses.
Moreover,
vaccination
elicited
antibodies
in
mice
neutralized
animal
human
alphacoronaviruses
betacoronaviruses
vitro
provided
degree
against
challenge
vivo.
Last,
history
Wuhan–based
vaccination,
further
broader
response
than
booster
Wuhan
suggesting
may
prevent
repertoire
focusing
caused
repeated
homologous
These
data
establish
protective
value
an
vaccine
support
notion
better
prepare
immune
system
respond
changing
nature
S1
subunit
concern,
as
well
future
zoonoses.
Science,
Год журнала:
2024,
Номер
385(6710), С. 757 - 765
Опубликована: Авг. 15, 2024
The
severe
acute
respiratory
syndrome
coronavirus
2
(SARS-CoV-2)
spike
protein
binds
the
receptor
angiotensin
converting
enzyme
(ACE2)
and
drives
virus-host
membrane
fusion
through
refolding
of
its
S2
domain.
Whereas
S1
domain
contains
high
sequence
variability,
is
conserved
a
promising
pan-betacoronavirus
vaccine
target.
We
applied
cryo-electron
tomography
to
capture
intermediates
understand
inhibition
by
antibodies
stem-helix.
Subtomogram
averaging
revealed
ACE2
dimers
cross-linking
spikes
before
transitioning
into
intermediates,
which
were
captured
at
various
stages
refolding.
Pan-betacoronavirus
neutralizing
targeting
stem-helix
bound
inhibited
prehairpin
intermediates.
Combined
with
molecular
dynamics
simulations,
these
structures
elucidate
process
SARS-CoV-2
entry
reveal
how
S2-targeting
neutralize
infectivity
arresting
The
outcome
of
infection
is
dependent
on
the
ability
viruses
to
manipulate
infected
cell
evade
immunity,
and
immune
response
overcome
this
evasion.
Understanding
process
key
understanding
pathogenesis,
genetic
risk
factors,
both
natural
vaccine-induced
immunity.
SARS-CoV-2
antagonises
innate
interferon
response,
but
whether
it
manipulates
cellular
immunity
unclear.
An
unbiased
proteomic
analysis
determined
how
surface
protein
expression
altered
SARS-CoV-2-infected
lung
epithelial
cells,
showing
downregulation
activating
NK
ligands
B7-H6,
MICA,
ULBP2,
Nectin1,
with
minimal
effects
MHC-I.
This
occurred
at
level
synthesis,
could
be
mediated
by
Nsp1
Nsp14,
correlated
a
reduction
in
activation.
identifies
novel
mechanism
which
host-shutoff
Later
disease
process,
strong
antibody-dependent
activation
(ADNKA)
developed.
These
responses
were
sustained
for
least
6
months
most
patients,
led
high
levels
pro-inflammatory
cytokine
production.
Depletion
spike-specific
antibodies
confirmed
their
dominant
role
neutralisation,
these
played
only
minor
ADNKA
compared
other
proteins,
including
ORF3a,
Membrane,
Nucleocapsid.
In
contrast,
induced
following
vaccination
was
focussed
solely
spike,
weaker
than
infection,
not
boosted
second
dose.
insights
have
important
implications
progression,
vaccine
efficacy,
design.
SARS-CoV-2
encodes
four
structural
proteins
incorporated
into
virions,
spike
(S),
envelope
(E),
nucleocapsid
(N),
and
membrane
(M).
M
plays
an
essential
role
in
viral
assembly
by
organizing
other
through
physical
interactions
directing
them
to
sites
of
budding.
As
the
most
abundant
protein
a
target
patient
antibodies,
is
compelling
for
vaccines
therapeutics.
Still,
structure
molecular
basis
its
virion
formation
are
unknown.
Here,
we
present
cryo-EM
lipid
nanodiscs
3.5
Å
resolution.
forms
50
kDa
homodimer
that
structurally
related
ORF3a
viroporin,
suggesting
shared
ancestral
origin.
Structural
comparisons
reveal
how
intersubunit
gaps
create
small,
enclosed
pocket
large
open
cavity
ORF3a,
consistent
with
ion
channel
activity,
respectively.
displays
strikingly
electropositive
cytosolic
surface
may
be
important
N,
S,
RNA.
Molecular
dynamics
simulations
show
high
degree
rigidity
simple
bilayer
support
homodimers
scaffolding
assembly.
Together,
these
results
provide
insight
roles
coronavirus
structure.
Emerging Microbes & Infections,
Год журнала:
2022,
Номер
11(1), С. 1037 - 1048
Опубликована: Март 23, 2022
The
coronavirus
SARS-CoV-2
is
the
causative
agent
for
disease
COVID-19.
To
capture
IgA,
IgG,
and
IgM
antibody
response
of
patients
infected
with
at
individual
epitope
resolution,
we
constructed
planar
microarrays
648
overlapping
peptides
that
cover
four
major
structural
proteins
S(pike),
N(ucleocapsid),
M(embrane),
E(nvelope).
arrays
were
incubated
sera
67
positive
22
negative
control
samples.
Specific
responses
to
detectable,
nine
associated
a
more
severe
course
disease.
A
random
forest
model
disclosed
binding
21
peptides,
mostly
localized
in
S
protein,
was
higher
neutralization
values
cellular
anti-SARS-CoV-2
assays.
For
antibodies
addressing
N-terminus
M,
or
close
fusion
region
S,
protective
effects
proven
by
depletion
study
pinpoints
unusual
viral
epitopes
might
be
suited
as
vaccine
candidates.
Proceedings of the National Academy of Sciences,
Год журнала:
2022,
Номер
119(31)
Опубликована: Июль 15, 2022
Camelid
single-domain
antibodies,
also
known
as
nanobodies,
can
be
readily
isolated
from
naïve
libraries
for
specific
targets
but
often
bind
too
weakly
to
their
immediately
useful.
Laboratory-based
genetic
engineering
methods
enhance
affinity,
termed
maturation,
deliver
useful
reagents
different
areas
of
biology
and
potentially
medicine.
Using
the
receptor
binding
domain
(RBD)
severe
acute
respiratory
syndrome
coronavirus
2
(SARS-CoV-2)
spike
protein
a
library,
we
generated
closely
related
nanobodies
with
micromolar
nanomolar
affinities.
By
analyzing
structure–activity
relationship
using
X-ray
crystallography,
cryoelectron
microscopy,
biophysical
methods,
observed
that
higher
conformational
entropy
losses
in
formation
protein–nanobody
complex
are
associated
tighter
binding.
To
investigate
this,
structural
ensembles
complexes
electron
microscopy
maps
correlated
fluctuations
affinity.
This
insight
guided
nanobody
improved
affinity
protein.